title: Isolation of Lactobacillus salivarius 1077 (NRRL B-50053) and characterization of its bacteriocin, including the antimicrobial activity spectrum.
abstract: Lactobacillus salivarius 1077 (NRRL B-50053) was isolated from poultry intestinal materials, and in vitro anti-Campylobacter jejuni activity was demonstrated. The isolate was then used for bacteriocin production and its enrichment. The protein content of the cell-free supernatant from the spent medium was precipitated by ammonium sulfate and dialyzed to produce the crude antimicrobial preparation. A typical bacteriocin-like response of sensitivity to proteolytic enzymes and resistance to lysozyme, lipase, and 100°C was observed with this preparation. The polypeptide was further purified by gel filtration, ion-exchange, and hydrophobic-interaction chromatography. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), Edman degradation, and isoelectrofocusing were used to characterize its 3,454-Da molecular mass, the amino acid sequence of its 37 residue components, and the isoelectric point of pI 9.1 of the bacteriocin. Bacteriocin L-1077 contained the class IIa bacteriocin signature N-terminal sequence YGNGV. MICs of bacteriocin L-1077 against 33 bacterial isolates (both Gram negative and Gram positive) ranged from 0.09 to 1.5 μg/ml. Subsequently, the therapeutic benefit of bacteriocin L-1077 was demonstrated in market-age (40- to 43-day-old) broiler chickens colonized with both C. jejuni and Salmonella enterica serovar Enteritidis. Compared with untreated control birds, both C. jejuni and S. Enteritidis counts in colonized ceca were diminished by >4 log(10) and S. Enteritidis counts in both the liver and the spleen of treated birds were reduced by 6 to 8 log(10)/g compared with those in the nontreated control birds. Bacteriocin L-1077 appears to hold promise in controlling C. jejuni/S. Enteritidis among commercial broiler chickens.
authors.affiliation: State Research Center for Applied Microbiology and Biotechnology, Obolensk, Russian Federation.
authors.forename: Edward A
authors.initials: EA
authors.lastname: Svetoch
authors.forename: Boris V
authors.initials: BV
authors.lastname: Eruslanov
authors.forename: Vladimir P
authors.initials: VP
authors.lastname: Levchuk
authors.forename: Vladimir V
authors.initials: VV
authors.lastname: Perelygin
authors.forename: Evgeny V
authors.initials: EV
authors.lastname: Mitsevich
authors.forename: Irina P
authors.initials: IP
authors.lastname: Mitsevich
authors.forename: Juri
authors.initials: J
authors.lastname: Stepanshin
authors.forename: Ivan
authors.initials: I
authors.lastname: Dyatlov
authors.forename: Bruce S
authors.initials: BS
authors.lastname: Seal
authors.forename: Norman J
authors.initials: NJ
authors.lastname: Stern
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents; D001430:Bacteriocins / Q000032:analysis / Q000494:pharmacology* / Q000627:therapeutic use; D016123:Campylobacter jejuni / Q000187:drug effects; D002645:Chickens / Q000382:microbiology; D004751:Enteritis / Q000188:drug therapy* / Q000382:microbiology; D005516:Food Microbiology; D007525:Isoelectric Focusing; D007778:Lactobacillus / Q000145:classification / Q000302:isolation & purification* / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D019779:Salmonella enterica / Q000187:drug effects; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000900:Anti-Bacterial Agents; D001430:Bacteriocins